Gupta R K, Siber G R
Massachusetts Public Health Biologic Laboratories, State Laboratory Institute, Jamaica Plain 02130.
Biologicals. 1994 Mar;22(1):53-63. doi: 10.1006/biol.1994.1008.
The adjuvant activity of three adjuvants, aluminium phosphate (AIPO4), calcium phosphate (CaHPO4) and stearyl tyrosine for tetanus toxoid (TT) were compared to soluble TT in mice at a dose of 0.5 Lf (1/10th of the single human dose) and in guinea-pigs at a dose of 7.5 Lf (1.5 times the single human dose). Three TT preparations varying in purity were used: (1) ammonium sulphate precipitated formalin detoxified tetanus toxin (AS-TT); (2) AS-TT ultrafiltered to remove low molecular weight peptides (UF-TT): and (3) chromatographically purified tetanus toxin subsequently detoxified with formalin (CP-TT). After primary immunization of mice, AIPO4 absorbed TTs induced higher toxin-neutralizing and IgG (by ELISA) antibodies than CaHPO4, stearyl tyrosine adsorbed or soluble TT preparations, but this difference was no longer present after secondary immunization. TT preparations of varying purities showed similar antibody responses after primary and secondary immunizations when adsorbed on each adjuvant. CP-TT preparation showed the highest neutralizing antibody level amongst soluble preparations after the first dose. All the preparations induced mainly IgG1 antibodies. However, stearyl tyrosine adsorbed TT induced relatively higher IgG2a and IgG2b responses than AIPO4, CaHPO4 adsorbed or soluble TTs particularly after booster injection. No preparation induced detectable IgG3 or IgM antibodies. AIPO4 adsorbed preparations induced higher IgE antibodies than CaHPO4 and stearyl tyrosine adsorbed vaccines. Among the soluble preparations, CP-TT induced lower anti-TT IgE antibodies than standard AS-TT. All these preparations were also tested in the US potency test for adsorbed TT in guinea-pigs. While all the preparations passed this test, AIPO4 adsorbed TT preparations induced higher neutralizing and IgG antibodies than CaHPO4 and stearyl tyrosine adsorbed or soluble TT preparations. In these animal models, purified TT was a strong immunogen and traditional AIPO4 adjuvant gave the highest antibody responses.
将三种佐剂，即磷酸铝（AIPO4）、磷酸钙（CaHPO4）和硬脂酰酪氨酸与破伤风类毒素（TT）的佐剂活性，在小鼠中以0.5Lf（单人剂量的1/10）的剂量以及在豚鼠中以7.5Lf（单人剂量的1.5倍）的剂量与可溶性TT进行了比较。使用了三种纯度不同的TT制剂：（1）硫酸铵沉淀福尔马林解毒破伤风毒素（AS-TT）；（2）经超滤去除低分子量肽的AS-TT（UF-TT）；以及（3）经色谱纯化后用福尔马林解毒的破伤风毒素（CP-TT）。在小鼠初次免疫后，AIPO4吸附的TTs诱导的毒素中和抗体和IgG（通过ELISA法）抗体高于CaHPO4、硬脂酰酪氨酸吸附的或可溶性TT制剂，但二次免疫后这种差异不再存在。不同纯度的TT制剂在吸附于每种佐剂后，初次和二次免疫后的抗体反应相似。CP-TT制剂在首剂后在可溶性制剂中显示出最高的中和抗体水平。所有制剂主要诱导IgG1抗体。然而，硬脂酰酪氨酸吸附的TT诱导的IgG2a和IgG2b反应相对高于AIPO4、CaHPO4吸附的或可溶性TTs，特别是在加强注射后。没有制剂诱导出可检测到的IgG3或IgM抗体。AIPO4吸附的制剂诱导的IgE抗体高于CaHPO4和硬脂酰酪氨酸吸附的疫苗。在可溶性制剂中，CP-TT诱导的抗TT IgE抗体低于标准AS-TT。所有这些制剂也在豚鼠吸附TT的美国效力试验中进行了测试。虽然所有制剂都通过了该试验，但AIPO4吸附的TT制剂诱导的中和抗体和IgG抗体高于CaHPO4、硬脂酰酪氨酸吸附的或可溶性TT制剂。在这些动物模型中，纯化的TT是一种强免疫原，传统的AIPO4佐剂产生的抗体反应最高。
